The operation of Dendritic Cell (DC) Core will be the responsibility of ImmunoSite, Inc., a private sector laboratory established as a joint venture between the University of Pittsburgh Medical Center (UPMC) and a local investment group. ImmunoSite, Inc. has a subcontract with the only GMP facility in Pittsburgh, i.e., Immunologic Monitoring and Cellular Products Laboratory (IMCPL) at the UPMC, for culture of clinical-grade DC. Under the terms of the existing subcontract, the IMCPL, functioning as a GMP facility, will perform all the work necessary for supporting the clinical tdals, which propose to use apoptotic cells infected with autologous HIV and fed to polarized DC 1 as a vaccine for HIV subjects concurrently undergoing HAART therapy. The objectives of Core D are: (1) to perform pre-clinical studies designed to select the ex vivo conditions for DC polarization and to optimize the use of a self-contained Aastrom Replicell System for culture of human DC and their infection with autologous HIV in years land 2;(2) to apply these optimized methodologies to a large-scale clinical product generation for clinical trials planned as part of project 3 during years 3 and 4;and (3) to serially monitor HIV-specific responses in subjects treated with the vaccine, using the state-of-the-art single cell methods for frequency measurements of HIV-specific T-cells in the peripheral circulation. Specifically, Core D will culture and phenotypically/functionally characterize DC for therapy and prepare vaccines by feeding these DC with apoptotic lymphocytes infected with autologous HIV. Core D will be responsible for culture and expansion of HIV-containing autologous T lymphocytes, HIV inactivation prior to the uptake of apoptoUc cells by DC and for the quality as well as sterility of the DC-cell products destined for therapy. It will also procure and process peripheral blood cells or body fluids harvested in the course of the clinical protocols, assay these specimens, generate immune data and distribute remaining cells to the participating investigators. The Core will ensure that all cellular products it generates and samples it collects are accompanied by appropriate documentation that will permit linking laboratory analyses with clinical results.